RAD-Seq of Rana muscosa: SJ population
MboI (GATC/CTAG) and PstI (CTGCAG/GACGTC) were used for library preparation. DNA samples were digested for 4 hours with MboI/PstI and indexes were ligated for 8 hours at 16C. An initial 15 cycle PCR reaction was performed to add a biotinylated primer and second index to the pools, and samples were purified on streptavidin beads. Final libraries were made by running 4 cycles of PCR on eluted DNA. Individual libraries were pooled at equimolar ratios, precipitated and size selected (280-500 bp) on Pippin Prep.